FACS-Service

Since 2014, the technical service for FACS flow-cytometry is also affiliated with the microscopy service group. The FACS technology is based on selective labelling of cells using specific antibodies conjugated with fluorescent dyes or expression of endogenous fluorophores.  In a flow-cytometer, the fluorescence signal is then exploited to count or sort individual cells bearing the fluorescence marker. Our institute currently hosts two flow-cytometers which are accessible for all members of the institute and which can be booked via the internal booking system:

 

Accuri C6 (BD):

  • Analytical flow-cytometer (easy-to-use, no cell sorting)
  • 473nm (blue) and 552nm (green) laser
  • 9 filter for simultaneous detection of e.g. GFP (or Alexa488) and mCherry (or Alexa546)

Aria II SORP (BD):

  • Cell-Sorter (allows cell sorting and single cell sorting)
  • Fixed configurations of laser and filter sets: Current available combinations are:
  • 458nm laser (darkblue for e.g. CFP) and one filter
  • 488nm laser (blue for e.g. GFP) with 6 filters
  • 561nm laser (green for e.g. mCherry) with 2 filters
  • 633nm laser (red for e.g. APC) with 2 filters

 As “sheath buffer”, sterile Coulter Isoton II buffer (Beckman) is routinely used. Samples can be analyszed at different temperatures using up to 4 tubes with variable size and 96-well plates, respectively.

Available nozzle sizes: 70, 85 (standard) and 100µm

Software: DIVA 6.1.3

More information on the instruments is available on the FACS twiki:

 

Prior to using the instruments, please contact Uta Marchfelder for planning your FACS experiments and for receiving proper instructions as how to use the devices. When planning and designing FACS experiments, it is strongly recommended to check the spectral properties as well as potential spectral overlaps of the desired fluorophores. For further advice, use e.g. the BD spectrumviewer or contact us:

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